Vaccine

Abstract

The present invention relates to Human Rhinovirus (HRV) Virus-Like Particles (VLPs) and methods of making HRV VLPs.

1 .- 2 . (canceled) 
     
     
         3 . A Virus-Like Particle (VLP), comprising human rhinovirus capsid proteins VP0, VP1 and VP 3. 
     
     
         4 . The VLP of  claim 3 , further comprising capsid protein VP 4. 
     
     
         5 . The VLP of  claim 3 , wherein all capsid proteins originate from the same seroptype of human rhinovirus. 
     
     
         6 . The VLP of  claim 3 , wherein the capsid proteins originate from two or more different serotypes of human rhinovirus. 
     
     
         7 . The VLP of  claim 3 , wherein one or more of the capsid proteins comprises at least one modification which increases the ability of the VLP to induce a cross-reactive immune response against multiple human rhinoviral serotypes, as compared to a VLP wherein said modification is not present, where said modification consists of insertion of at least one peptide from a rhinoviral capsid protein, said peptide capable of inducing a cross-reactive immune response against two or more human rhinoviral serotypes. 
     
     
         8 .- 10 . (canceled) 
     
     
         11 . The VLP of  claim 7 , wherein said peptide comprises a sequence selected from (a) amino acids 32-45 from VP1 SEQ ID NO:15, (b) amino acids 32-45 of VP1 SEQ ID NO:15 having 1-4 amino acid deletions at either end, (c) amino acids 32-45 of VP1 SEQ ID NO:15 having 1-2 amino acid substitutions or additions or deletions, (d) SEQ ID NO: 7, (e) SEQ ID NO:8, (f) SEQ ID NO:9, and (q) SEQ ID NO:10. 
     
     
         12 . (canceled) 
     
     
         13 . The VLP of  claim 7 , wherein said peptide comprises a sequence selected from (a) amino acids 1-16 from VP4 SEQ ID NO:18, (b) amino acids 1-16 of VP4 SEQ ID NO: 18 having 1-4 amino acid deletions at either end, (c) amino acids 1-16 of VP4 SEQ ID NO:18 having 1-2 amino acid substitutions or additions or deletions, (d) SEQ ID NO:11, (e) SEQ ID NO:12, and (f) SEQ ID NO:13. 
     
     
         14 . (canceled) 
     
     
         15 . The VLP of  claim 7 , wherein said peptide is selected from:
 (a) amino acids 147-162 of VP1 SEQ ID NO:15,   (b) amino acids 147-162 of VP1 SEQ ID NO:15, having 1-4 amino acid additions or deletions at either end,   (c) amino acids 147-162 of VP1 SEQ ID NO:15, having 1-2 amino acid substitutions or additions or deletions within the peptide sequence,   (d) amino acids 1-30 of VP4 SEQ ID NO:18,   (e) amino acids 1-30 of VP4 SEQ ID NO:18, having 1-4 amino acid additions or deletions at either end,   (f) amino acids 1-30 of VP4 SEQ ID NO:18, having 1-2 amino acid substitutions or additions or deletions within the peptide sequence,   (g) amino acids 1-24 of VP4 SEQ ID NO:18,   (h) amino acids 1-24 of VP4 SEQ ID NO:18, having 1-4 amino acid additions or deletions at either end,   (i) amino acids 1-24 of VP4 SEQ ID NO:18, having 1-2 amino acid substitutions or additions or deletions within the peptide sequence,   (j) amino acids 1-8 of VP1 SEQ ID NO:15,   (k) amino acids 1-8 of VP1 SEQ ID NO:15, having 1-4 amino acid additions or deletions at either end,   (l) amino acids 1-8 of VP1 SEQ ID NO:15, having 1-2 amino acid substitutions or additions or deletions within the peptide sequence,   (m) amino acids 277-283 of VP1 SEQ ID NO:15,   (n) amino acids 277-283 of VP1 SEQ ID NO:15, having 1-4 amino acid additions or deletions at either end,   (o) amino acids 277-283 of VP1 SEQ ID NO:15, having 1-2 amino acid substitutions or additions or deletions within the peptide sequence,   (p) amino acids 275-285 of VP1 SEQ ID NO:15,   (q) amino acids 275-285 of VP1 SEQ ID NO:15, having 1-4 amino acid additions or deletions at either end, and   (r) amino acids 275-285 of VP1 SEQ ID NO:15, having 1-2 amino acid substitutions or additions or deletions within the peptide sequence.   
     
     
         16 .- 25 . (canceled) 
     
     
         26 . The VLP of  claim 3 , wherein the VLP comprises at least one modification which results in attenuation of an immunodominant epitope. 
     
     
         27 . The VLP of  claim 26 , wherein VP0 comprises a modification selected from: (a) attenuation of the NIm-II site epitope, (b) modification of one or more residues in the regions corresponding to amino acids 131-175, 228-238 or 255-262 in VP2 SEQ ID NO:16, and (c) modification of a residue corresponding to residue 158, 159, 161 and/or 162 of VP2 SEQ ID NO:16. 
     
     
         28 . The VLP of  claim 26 , wherein VP1 comprises a modification selected from: (a) attenuation of the NIm-IA site epitope (b) attenuation of the NIm-IB site epitope (c) modification of one or more residues in the regions corresponding to amino acids 73-102, 129-149, 204-212 and 264-289 in VP1 SEQ ID NO:15, and (d) modification of a residue corresponding to residue 83, 85, 91, 95, 138 or 139 in VP1 SEQ ID NO:15. 
     
     
         29 . The VLP of  claim 26 , wherein VP2 comprises a modification selected from: (a) attenuation of the NIm-II site epitope, (b) modification of one or more residues in the regions corresponding to amino acids 131-175, 228-238 or 255-262 in VP2 SEQ ID NO:16, and (c) modification of a residue corresponding to residue 158, 159, 161 or 162 in VP2 SEQ ID NO:16. 
     
     
         30 . The VLP of  claim 26 , wherein VP3 comprises a modification selected from: (a) attenuation of the NIm-Ill site epitope, (b) modification of one or more residues in the regions corresponding to amino acids 54-95, 193-212 or 226-236 in VP3 of SEQ ID NO: 17, and (c) modification of a residue corresponding to residue 72, 75 or 78 in VP3 SEQ ID NO: 17. 
     
     
         31 . The VLP of  claim 4 , wherein the VLP comprises at least one modification in capsid protein VP4 which results in attenuation of an immunodominant epitope. 
     
     
         32 . The VLP of  claim 3 , wherein the VP0 protein has a sequence selected from the group consisting of (a) SEQ ID NO:14; (b) a sequence having more than 90% sequence identity to SEQ ID NO:14, (c) SEQ ID NO:14 having from 1-25 amino acid additions or deletions at either end, and (d) SEQ ID NO:14 having 1-50 amino acid substitutions or deletions within the sequence. 
     
     
         33 . (canceled) 
     
     
         34 . The VLP of  claim 3 , wherein the VP1 protein has a sequence selected from the group consisting of (a) SEQ ID NO:15, (b) a sequence having more than 90% sequence identity to SEQ ID NO:15, (c) SEQ ID NO:15 having from 1-25 amino acid additions or deletions at either end, and (d) SEQ ID NO:15 having 1-50 amino acid substitutions or deletions within the sequence. 
     
     
         35 . (canceled) 
     
     
         36 . The VLP of  claim 3 , wherein the VP2 protein has a sequence selected from the group consisting of (a) SEQ ID NO:16, (b) a sequence having more than 90% sequence identity to SEQ ID NO:16, (c) SEQ ID NO:16 having from 1-25 amino acid additions or deletions at either end, and (d) SEQ ID NO:16 having 1-50 amino acid substitutions or deletions within the sequence. 
     
     
         37 . (canceled) 
     
     
         38 . The VLP of  claim 3 , wherein the VP3 protein has a sequence selected from the group consisting of (a) SEQ ID NO: 17, (b) a sequence having more than 90% sequence identity to SEQ ID NO:17, (c) SEQ ID NO:17 having from 1-25 amino acid additions or deletions at either end, and (d) SEQ ID NO:17 having 1-50 amino acid substitutions or deletions within the sequence. 
     
     
         39 . (canceled) 
     
     
         40 . The VLP of  claim 3 , wherein the VP4 protein has a sequence selected from the group consisting of (a) SEQ ID NO:18, (b) a sequence having more than 90% sequence identity to HRV14 VP4 SEQ ID NO:18, (c) SEQ ID NO:18 having from 1-25 amino acid additions or deletions at either end, and (d) SEQ ID NO:18 having 1-50 amino acid substitutions or deletions within the sequence. 
     
     
         41 . (canceled) 
     
     
         42 . An immunogenic composition comprising the VLP of  claim 3  and a pharmaceutically acceptable diluent, excipient or carrier. 
     
     
         43 .- 48 . (canceled) 
     
     
         49 . A method for inducing an immune response in a human against human rhinovirus comprising administering to said human an immunogenic composition according to  claim 42 . 
     
     
         50 .- 53 . (canceled) 
     
     
         54 . A method for producing rhinoviral VLPs comprising the steps of:
 (a) producing constructs encoding capsid proteins required for the formation of a VLP, wherein each of the capsid proteins is encoded as a fusion protein with a chaperone protein, and   (b) expressing said constructs in a host cell, and   (c) lysing said host cells.   
     
     
         55 . (canceled) 
     
     
         56 . The method according to  claim 54 , wherein said chaperone protein is a Small Ubiquitin-related Modifier (SUMO) protein. 
     
     
         57 .- 67 . (canceled)